Extended hypoxia-mediated H₂ S production provides for long-term oxygen sensing

Numerous studies have shown that H₂ S serves as an acute oxygen sensor in a variety of cells. We hypothesize that H₂ S also serves in extended oxygen sensing.

Here, we compare the effects of extended exposure (24-48 hours) to varying O₂ tensions on H₂ S and polysulphide metabolism in human embryonic kidney (HEK 293), human adenocarcinomic alveolar basal epithelial (A549), human colon cancer (HTC116), bovine pulmonary artery smooth muscle, human umbilical-derived mesenchymal stromal (stem) cells and porcine tracheal epithelium (PTE) using sulphur-specific fluorophores and fluorometry or confocal microscopy.

All cells continuously produced H₂ S in 21% O₂ and H₂ S production was increased at lower O₂ tensions. Decreasing O₂ from 21% to 10%, 5% and 1% O₂ progressively increased H₂ S production in HEK293 cells and this was partially inhibited by a combination of inhibitors of H₂ S biosynthesis, aminooxyacetate, propargyl glycine and compound 3. Mitochondria appeared to be the source of much of this increase in HEK 293 cells. H₂ S production in all other cells and PTE increased when O₂ was lowered from 21% to 5% except for HTC116 cells where 1% O₂ was necessary to increase H₂ S, presumably reflecting the hypoxic environment in vivo. Polysulphides (H₂ S_n , where n = 2-7), the key signalling metabolite of H₂ S also appeared to increase in many cells although this was often masked by high endogenous polysulphide concentrations.

These results show that cellular H₂ S is increased during extended hypoxia and they suggest this is a continuously active O₂ -sensing mechanism in a variety of cells.